Buffered Charcoal Yeast Extract Differential Agar (DIFF/BCYE)

[所属分类:培养基配方] [发布时间:2020-3-18] [发布人:] [阅读次数:] [返回]

山东拓普生物工程有限公司

Shandong Tuopu Biol-Engineering Co.,Ltd

Buffered Charcoal Yeast Extract Differential Agar
(DIFF/BCYE)

培养基配方
Composition per 1014.0mL:
Agar ............................................................17.0g
ACES (2-[(2-amino-2-oxoethyl)-
amino]-ethane sulfonic acid) buffer .............................10.0g
Yeast extract....................................................10.0g
Charcoal, activated...............................................1.5g
Fe4(P2O7)3 ·9H2O................................................0.25g
Bromcresol Purple ...............................................0.01g
Bromthymol Blue .................................................0.01g
Antibiotic solution ............................................10.0mL
L -Cysteine·HCl·H 2 O solution ................................4.0mL
pH 6.9 ± 0.2 at 25°C
Antibiotic Solution:
Composition per 10.0mL:
Vancomycin .....................................................1.0mg
Polymyxin B...................................................50,000U
Preparation of Antibiotic Solution: Add components to distilled/
deionized water and bring volume to 10.0mL. Mix thoroughly. Filter
sterilize.
L -Cysteine·HCl·H2O Solution:
Composition per 10.0mL:
L- Cysteine·HCl·H2O.........................................1.0g
Preparation of  L -Cysteine·HCl·H2O Solution: Add 1.0g of  L- cys-
teine·HCl·H2O to distilled/deionized water and bring volume to
10.0mL. Mix thoroughly. Filter sterilize.
Preparation of Medium:  Add components, except  L -cysteine·HCl·H2O
solution and antibiotic solution, to distilled/deionized water and bring vol-
ume to 1.0L. Mix thoroughly. Adjust medium to pH 6.9 with 1N KOH.
Heat gently and bring to boil for 1 min. Autoclave for 15 min at 15 psi pres-
sure–121°C. Cool to 50°–55°C. Add 4.0mL of sterile L- cysteine·HCl·H2O
solution and 10.0mL of sterile antibiotic solution. Mix thoroughly. Pour
into sterile Petri dishes with constant agitation to keep charcoal in suspen-
sion.
Use:  For the isolation, cultivation, and maintenance of Legionella
pneumophila and other Legionella species from environmental and
clinical specimens. For the selective recovery of Legionella pneumo-
phila while reducing contaminating microorganisms from environ-
mental water samples.